LOCUS       sequence-231063-        6364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6364)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 6364)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CHL1 P492L
FEATURES             Location/Qualifiers
     source          1..6364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             2716..2742
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             2752..2769
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2849..3095
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3110..3523
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3530..3577
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3596..3970
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      4036..4283
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4052..4070)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(5696..6284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5776..5795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg aggaattcat gtctcttcct
      961 gaaactaaat ctgatgatat ccttcttgat gcttgggact tccaaggccg tcccgccgat
     1021 cgctcaaaaa ccggcggctg ggccagcgcc gccatgattc tttgtattga ggccgtggag
     1081 aggctgacga cgttaggtat cggagttaat ctggtgacgt atttgacggg aactatgcat
     1141 ttaggcaatg caactgcggc taacaccgtt accaatttcc tcggaacttc tttcatgctc
     1201 tgtctcctcg gtggcttcat cgccgatacc tttctcggca ggtacctaac gattgctata
     1261 ttcgccgcaa tccaagccac gggtgtttca atcttaactc tatcaacaat cataccggga
     1321 cttcgaccac caagatgcaa tccaacaacg tcgtctcact gcgaacaagc aagtggaata
     1381 caactgacgg tcctatactt agccttatac ctcaccgctc taggaacggg aggcgtgaag
     1441 gctagtgtct cgggtttcgg gtcggaccaa ttcgatgaga ccgaaccaaa agaacgatcg
     1501 aaaatgacat atttcttcaa ccgtttcttc ttttgtatca acgttggctc tcttttagct
     1561 gtgacggtcc ttgtctacgt acaagacgat gttggacgca aatggggcta tggaatttgc
     1621 gcgtttgcga tcgtgcttgc actcagcgtt ttcttggccg gaacaaaccg ctaccgtttc
     1681 aagaagttga tcggtagccc gatgacgcag gttgctgcgg ttatcgtggc ggcgtggagg
     1741 aataggaagc tcgagctgcc ggcagatccg tcctatctct acgatgtgga tgatattatt
     1801 gcggcggaag gttcgatgaa gggtaaacaa aagctgccac acactgaaca attccgttca
     1861 ttagataagg cagcaataag ggatcaggaa gcgggagtta cctcgaatgt attcaacaag
     1921 tggacactct caacactaac agatgttgag gaagtgaaac aaatcgtgcg aatgttacca
     1981 atttgggcaa catgcatcct cttctggacc gtccacgctc aattaacgac attatcagtc
     2041 gcacaatccg agacattgga ccgttccatc gggagcttcg agatccctcc agcatcgatg
     2101 gcagtcttct acgtcggtgg cctcctccta accaccgccg tctatgaccg cgtcgccatt
     2161 cgtctatgca aaaagctatt caactacccc catggtctaa gaccgcttca acggatcggt
     2221 ttggggcttt tcttcggatc aatggctatg gctgtggctg ctttggtcga gctcaaacgt
     2281 cttagaactg cacacgctca tggtccaaca gtcaaaacgc ttcctctagg gttttatcta
     2341 ctcatcccac aatatcttat tgtcggtatc ggcgaagcgt taatctacac aggacagtta
     2401 gatttcttct tgagagagtg ccttaaaggt atgaaaggga tgagcacggg tctattgttg
     2461 agcacattgg cattaggctt tttcttcagc tcggttctcg tgacaatcgt cgagaaattc
     2521 accgggaaag ctcatccatg gattgccgat gatctcaaca agggccgtct ttacaatttc
     2581 tactggcttg tggccgtact tgttgccttg aacttcctca ttttcctagt tttctccaag
     2641 tggtacgttt acaaggaaaa aagactagct gaggtgggga ttgagttgga tgatgagccg
     2701 agtattccaa tgggtcatca tcatcatcat catcatcatc attaagtcga ccatcatcat
     2761 catcatcatt gagtttgtag ccttagacat gactgttcct cagttcaagt tgggcactta
     2821 cgagaagacc ggtcttgcta gattctaatc aagaggatgt cagaatgcca tttgcctgag
     2881 agatgcaggc ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt
     2941 ttgtcatttt gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagctgatg
     3001 aatatcttgt ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc
     3061 ccactcctct tcagagtaca gaagattaag tgagaccttc gtttgtgcgg atcccccaca
     3121 caccatagct tcaaaatgtt tctactcctt ttttactctt ccagattttc tcggactccg
     3181 cgcatcgccg taccacttca aaacacccaa gcacagcata ctaaatttcc cctctttctt
     3241 cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa gagaccgcct
     3301 cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc tttttcttga
     3361 aaattttttt ttttgatttt tttctctttc gatgacctcc cattgatatt taagttaata
     3421 aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt
     3481 acttcttgct cattagaaag aaagcatagc aatctaatct aagggcggtg ttgacaatta
     3541 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     3601 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     3661 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     3721 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     3781 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     3841 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     3901 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     3961 gcaggactga cacgtccgac gcggcccgac gggtccgagg cctcggagat ccgtccccct
     4021 tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     4081 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     4141 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     4201 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     4261 gacgctcgaa ggctttaatt tgcaagctgg agaccaacat gtgagcaaaa ggccagcaaa
     4321 aggccaggaa ccgtaaaaag gccgcgttgc tctgagagat cccctcataa tttccccaaa
     4381 acgtaaccat gtgtgaatag attttgagta agcagggttg cagccacgag tgagtcttcc
     4441 cttgttattg tgtagccaga atgccgcaaa acttccatgc ctaagcgaac tgttgagagt
     4501 acgtttcgat ttctgactgt gttagcctgg aagtgcttgt cccaaccttg tttctgagca
     4561 tgaacgcccg caagccaaca tgttagttga agcatcaggg cgattagcag catgatatca
     4621 aaacgctctg agctgctcgt tcggctatgg cgtaggccta gtccgtaggc aggacttttc
     4681 aagtctcgga aggtttcttc aatctgcatt cgcttcgaat agatattaac aagttgtttg
     4741 ggtgttcgaa tttcaacagg taagttagtt gctagaatcc atggctcctt tgccgacgct
     4801 gagtagattt taggtgacgg gtggtgacaa tgagtccgtg tcgagcgctg attttttcgg
     4861 cctttagagc gagatttata caatagaatt tggcatgaga ttgggttgct tttagtcagc
     4921 ctcttatagc ctaaagtctt tgagtgacta gatgacatat catgtaagtt gctgataggt
     4981 ttccagtttt ccgctcctag gtctgcatat tgtacttttc ctcttactcg acttaaccag
     5041 taccaaccca gcttctcaac ggatttatac catggcactt taaagccagc atcactgaca
     5101 atgagcggtg tggtgttact cggtagaatg ctcgcaaggt cggctagaaa ttggtcatga
     5161 gctttctttg aacattgctc tgaaagcggg aacgctttct cataaagagt aacagaacga
     5221 ccgtgtagtg cgactgaagc tcgcaatacc ataagccgtt tttgctcacg gatatcagac
     5281 cagtcaacaa gtacaatggg catcgtattg cccgaacaga taaagctagc atgccaacgg
     5341 tatacagcga gtcgctcttt gtggaggtga cgattaccta acaatcggtc gattcgtttg
     5401 atgttatgtt ttgttctcgc tttggttggc aggttacggc caagttcggt aagagtgaga
     5461 gttttacagt caagtaaggc gtggcaagcc aacgttaagc tgttgagtcg ttttaagtgt
     5521 aattcggggc agaattggta aagagagtcg tgtaaaatat cgagttcgca cattttgttg
     5581 tctgattatt gatttttggc gaaaccattt gatcatatga caagatgtgt atctacctta
     5641 acttaatgat tttgataaaa atcattaggg gattcatcag cgcgttgctg gcgtttttcc
     5701 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5761 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5821 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5881 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5941 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6001 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6061 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6361 gatc
//
